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In Vitro Cellular & Developmental Biology. Animal

Frank Simione, Tim Sharp
Successfully recovering stored cryopreserved cells requires attention to two major factors, i.e., storage temperature and proper handling of the frozen samples. These key considerations are in turn impacted by how the inventory system is constructed, the frequency and process for retrieving specimens, as well as how the material is secured. In addition, shipping biological materials requires attention to the type of material transported, adherence to regulatory requirements, packaging materials and proper assembly, labeling, and engaging reputable carriers...
December 13, 2017: In Vitro Cellular & Developmental Biology. Animal
Charlotte Pushparajan, Juan Daniel Claus, Sean D G Marshall, Gabriel Visnovsky
The DSIR-HA-1179 coleopteran cell line has been identified as a susceptible and permissive host for the in vitro replication of the Oryctes nudivirus, which can be used as a biopesticide against the coconut rhinoceros beetle, pest of palms. The major challenge to in vitro large-scale Oryctes nudivirus production is ensuring process economy. This rests, among other requisites, on the use of low-cost culture media tailored to the nutritional and metabolic needs of the cell line, both in uninfected and infected cultures...
December 1, 2017: In Vitro Cellular & Developmental Biology. Animal
Marimu Sakumoto, Rieko Oyama, Mami Takahashi, Yoko Takai, Fusako Kito, Kumiko Shiozawa, Zhiwei Qiao, Makoto Endo, Akihiko Yoshida, Akira Kawai, Tadashi Kondo
Clear cell sarcoma (CCS) is an aggressive mesenchymal malignancy characterized by the unique chimeric EWS-ATF1 fusion gene. Patient-derived cancer models are essential tools for the understanding of tumorigenesis and the development of anti-cancer drugs; however, only a limited number of CCS cell lines exist. The objective of this study was to establish patient-derived CCS models. We established patient-derived CCS models from a 43-yr-old female patient. We prepared the patient-derived xenografts (PDXs) from tumor tissues obtained through biopsy or surgery and isolated stable cell lines from PDXs and the original tumor tissue...
December 1, 2017: In Vitro Cellular & Developmental Biology. Animal
Buwei Liu, Yu Shi, Hongbing He, Mingcheng Cai, Wudian Xiao, Xue Yang, Shiyi Chen, Xianbo Jia, Jie Wang, Songjia Lai
MicroRNAs (miRNAs) are a class of small non-coding RNA molecules, which play important roles in animals by targeting mRNA transcripts for translational repression. Many recent studies have shown that miRNAs are involved in the control of muscle development. In this study, the expression levels of miR-221 in different tissues and during rabbit skeletal muscle satellite cells (SMSCs) differentiation were detected. Gene ontology term enrichment was used to predict the potential biological roles of miR-221. A synthetic miR-221 mimic and a miR-221 inhibitor were used to investigate the functions of miR-221 during SMSCs proliferation and differentiation to further verify the functions of miR-221 in muscle development...
December 1, 2017: In Vitro Cellular & Developmental Biology. Animal
Sandhya Noronha, Lauren A C Alt, Taylor E Scimeca, Omran Zarou, Justyna Obrzut, Brian Zanotti, Elizabeth A Hayward, Akhil Pillai, Shubha Mathur, Joseph Rojas, Ribhi Salamah, Nalini Chandar, Michael J Fay
Liposarcoma is a malignant soft tissue tumor that originates from adipose tissue and is one of the most frequently diagnosed soft tissue sarcomas in humans. There is great interest in identifying novel chemotherapeutic options for treating liposarcoma based upon molecular alterations in the cancer cells. The Aurora kinases have been identified as promising chemotherapeutic targets based on their altered expression in many human cancers and cellular roles in mitosis and cytokinesis. In this study, we investigated the effects of an Aurora kinase A inhibitor (MK-5108), an Aurora kinase B inhibitor (AZD1152-HQPA), and a pan-Aurora kinase inhibitor (AMG 900) on undifferentiated SW-872 and well-differentiated 93T449 human liposarcoma cells...
December 1, 2017: In Vitro Cellular & Developmental Biology. Animal
Guohong Chen, Zhiteng Li, Shuai Su, Guobin Chang, Lingling Qiu, Pengfei Zhu, Yang Zhang, Qi Xu
Avian leukemia subgroup J (ALV-J) is one of the most detrimental neoplastic diseases in poultry production. However, the differences between somatic cells and immune cells post-infection remain poorly understood. The aim of our study was to detect the different responses in chicken to infection with ALV-J in different cell lines. In this study, we detected transcriptome expression changes during infection with ALV-J in chicken embryo fibroblast (CEF) and HD11 cell lines. RNA-Seq was used to determine the expression levels of mRNA transcripts from the two cell types after infection with ALV-J at 1, 4, and 7 dpi, and gene ontology analyses were used to cluster differentially expressed genes into pathways...
December 1, 2017: In Vitro Cellular & Developmental Biology. Animal
Joseph S Zakhari, Jacob Zabonick, Brian Gettler, Stuart K Williams
Adipose-derived stromal vascular fraction (SVF) is a heterogeneous cell source that contains endothelial cells, pericytes, smooth muscle cells, stem cells, and other accessory immune and stromal cells. The SVF cell population has been shown to support vasculogenesis in vitro as well vascular maturation in vivo. Matrigel, an extracellular matrix (ECM) mixture has been utilized in vitro to evaluate tube formation of purified endothelial cell systems. We have developed an in vitro system that utilizes freshly isolated SVF and ECM molecules both in pure form (fibrin, laminin, collagen) as well as premixed form (Matrigel) to evaluate endothelial tip cell formation, endothelial stalk elongation, and early stages of branching and inosculation...
December 1, 2017: In Vitro Cellular & Developmental Biology. Animal
Raymond W Nims, Yvonne Reid
Experiments using cell cultures are only valid to the extent that the cell culture is a true model system for the biological system being investigated. To assure that a cell line is and remains an appropriate biological model, its identity, purity, ploidy, and phenotype must be maintained. These characteristics comprise and determine the authenticity of a cell line. Routine monitoring of the cell line through microscopic examination of morphology can help to determine authenticity, as can the determination of phenotypic status...
December 1, 2017: In Vitro Cellular & Developmental Biology. Animal
Raymond W Nims, Paul J Price
A variety of biological and chemical contaminants can adversely impact cells in culture, ranging from outright destruction of the culture, mutation, phenotypic changes to relatively minor changes in morphology, or growth rate. There are various approaches to detecting and mitigating the risk of biological or microbial contaminants in cell cultures, and these are discussed in this article. Chemical contaminants typically arise from improper handling or sourcing of cell culture reagents, glassware, or other types of consumables...
December 1, 2017: In Vitro Cellular & Developmental Biology. Animal
Yongli Yue, Lichun Zhang, Xiang Zhang, Xueling Li, Haiquan Yu
Adipose-derived mesenchymal stem cells (ADSCs) are useful cell model to study adipogenesis and energy metabolism. However, the biological characteristics of bovine ADSCs (bADSCs) remain unclear. This study aimed to isolate and identify bADSCs and further investigate fatty acid (FA)-related gene expression and composition of FAs during adipogenesis. The growth curve showed the bADSCs of P5 cells had rapid proliferation superior to P10-P50. The colony formation assay showed colony number of P5 cells was higher than that of P50 cells (51...
November 30, 2017: In Vitro Cellular & Developmental Biology. Animal
Mona Nazemi Moghaddam, Razieh Jalal, Zohreh Zeraatkar
Anticancer role of oxindole compounds is well documented. Here, we synthesized new derivatives of 3-hydroxy-2-oxindole functionalized at position 3 (1a-f) which are expected to have antiproliferative activity in cancer cells. Human prostate cancer cell line (DU145) was treated with the synthesized derivatives at 40-μM concentration for 24, 48, and 72 h. Compounds 1-ethyl-3-hydroxy-1,1',3,3'-tetrahydro-2H,2'H-3,3'-biindole-2,2'-dione (1d), 5-bromo-1-ethyl-3-hydroxy-1,1',3,3'-2H,2'H-3,3'-biindole-2,2'-dione (1e), and 5-chloro-1-ethyl-3-hydroxy-1,1',3,3'-tetrahydro-2H,2'H-3,3'-biindole-2,2'-dione (1f) were found to significantly reduce DU145 cell viability at 48 and 72 h whereas no significant changes were observed up to 24 h...
November 9, 2017: In Vitro Cellular & Developmental Biology. Animal
Toshihiro Sera, Tateki Sumii, Ryosuke Fujita, Susumu Kudo
When the liver is damaged, hepatic stellate cells (HSCs) can change into an activated, highly migratory state. The migration of HSCs may be affected by shear stress due not only to sinusoidal flow but also by the flow in the space of Disse because this space is filled with blood plasma. In this study, we evaluated the effects of shear stress on HSC migration in a scratch-wound assay with a parallel flow chamber. At regions upstream of the wound area, the migration was inhibited by 0.6 Pa and promoted by 2...
November 9, 2017: In Vitro Cellular & Developmental Biology. Animal
John M Baust, Lia H Campbell, John W Harbell
Long-term storage of cell stocks insures that cells are available for use whenever needed. Cryopreservation of cells is the method of choice for preservation of important or rare cell stocks. There are several factors to consider when establishing a protocol for freezing, thawing, and recovery of cells after storage. These parameters may include cell concentration, cryoprotectant choice and concentration, and thawing rate among others. Further, the assessment of cell viability and/or function prior to and following cryopreservation is imperative in order to accurately determine downstream utility as well for optimizing the cryopreservation process...
November 2, 2017: In Vitro Cellular & Developmental Biology. Animal
Li Chen, Liping Liu, Yinghui Li, Jing Gao
Considering that chemotherapy resistance is vital to the progression of cervical carcinoma, emerging researchers are focused on developing anti-tumor drugs to assist the treatment efficiency of chemotherapy. Melatonin has anti-tumor activity via several mechanisms including its anti-proliferative and pro-apoptotic effects as well as its potent pro-oxidant action in tumor cells. Therefore, melatonin may be useful for the treatment of tumors in association with chemotherapy drugs. Here, we studied the effect and mechanism of melatonin on HeLa cells apoptosis under cisplatin (CIS) treatment, particularly focusing on the caspase-9-related apoptosis pathway and mitophagy-mediated anti-apoptotic mechanism...
October 25, 2017: In Vitro Cellular & Developmental Biology. Animal
Sophia R Bloch, John J Kim, Phuc H Pham, Peter V Hodson, Lucy E J Lee, Niels C Bols
The effect of selenium deprivation and addition on the American eel brain endothelial cell line (eelB) was studied in three exposure media: complete growth medium (L15/FBS), serum-free medium (L15), and minimal medium (L15/ex). L15/ex contains only galactose and pyruvate and allowed the deprivation of selenium on cells to be studied. In L15/ex, without any obvious source of selenium, eelB cells survived for at least 7 d, formed capillary-like structures (CLS) on Matrigel, and migrated to heal wounds. Three selenium compounds were added to cultures: selenite, selenate, and selenomethionine (SeMet)...
September 22, 2017: In Vitro Cellular & Developmental Biology. Animal
Yvonne A Reid
One of the first considerations in using an existing cell line or establishing a new a cell line is the detailed proactive planning of all phases of the cell line management. It is necessary to have a well-trained practitioner in best practices in cell culture who has experience in receiving a new cell line into the laboratory, the correct and appropriate use of a cell line name, the preparation of cell banks, microscopic observation of cells in culture, growth optimization, cell count, cell subcultivation, as well as detailed protocols on how to expand and store cells...
October 2017: In Vitro Cellular & Developmental Biology. Animal
Alice P Christensen, Emeline Peyrache, Heidy Kaune, Suzannah A Williams
Basement membranes are found in every organ of the body. They provide structure and a selective filter for molecules. The ovary is no different with the follicular basal lamina (FBL) separating the granulosa and theca cells, facilitating regulation of the changing follicular environment providing appropriate conditions for the developing oocyte. The FBL is modified in C1galt1 Mutant mice (C1galt1 (FF):ZP3Cre) resulting from oocyte-specific deletion of C1galt1. Changes in the FBL lead to follicles joining to generate multiple-oocyte follicles (MOFs); where two or more oocytes are contained within a single follicle...
October 2017: In Vitro Cellular & Developmental Biology. Animal
Samaneh Kamalipooya, Parviz Abdolmaleki, Zahra Salemi, Fatemeh Javani Jouni, Jaber Zafari, Homa Soleimani
In this study, we reported the effects of simultaneous application of static magnetic field (SMF) and cisplatin as an anticancer drug on the oxidative stress in human cervical cancer (HeLa) cell line and normal skin fibroblast cells (Hu02). The cells were exposed to different SMF intensities (7, 10, and 15 mT) for 24 and 48 h. IC50 concentrations of cisplatin were obtained by MTT assay. The cytotoxic effects of combined treatment were studied by measuring the intracellular reactive oxygen species content using flow cytometric method and estimation of membrane lipid peroxidation by spectrophotometry...
October 2017: In Vitro Cellular & Developmental Biology. Animal
Saoko Tachikawa, Toshinobu Nishimura, Hiromitsu Nakauchi, Kiyoshi Ohnuma
Thalidomide, which was formerly available commercially to control the symptoms of morning sickness, is a strong teratogen that causes fetal abnormalities. However, the mechanism of thalidomide teratogenicity is not fully understood; thalidomide toxicity is not apparent in rodents, and the use of human embryos is ethically and technically untenable. In this study, we designed an experimental system featuring human-induced pluripotent stem cells (hiPSCs) to investigate the effects of thalidomide. These cells exhibit the same characteristics as those of epiblasts originating from implanted fertilized ova, which give rise to the fetus...
October 2017: In Vitro Cellular & Developmental Biology. Animal
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